Molecular Biology Core Laboratory

分子生物学核心实验室由罗素博士领导。他的日常工作将得到乔纳森·科恩博士的协助，他是高通量DNA测序方面的专家。这个核心实验室为蛋白质的丙烯酰胺凝胶电泳、DNA测序、通过寡核苷酸微阵列杂交和实时聚合酶链式反应(PCRS)分析基因表达、寡核苷酸获取、基因组DMA和RNA分离以及本计划项目中使用的细菌菌株、质粒和纯化蛋白的维护和储存提供支持。四名经验丰富的技术人员，凯文·安德森(100%时间)、艾米丽·布朗(100%)、杰弗里·科米尔(100%)和斯科特·克拉克(25%)将履行与该核心相关的职责。该实验室设在分子遗传学系内。 对于蛋白质的分析，有30个垂直电泳仪和12个多出口电源可供选择。SDS聚丙烯酰胺凝胶要么制备(凝胶的25%)，要么购买预先浇注的(凝胶的75%)，由一名技术员(艾米丽·布朗)操作。在电泳之后，从事不同研究项目的个人研究人员处理凝胶，用于放射自显影、免疫印迹或蛋白质测序。 一个装有柯尼卡自动X射线显影剂的暗室被用来处理所有的放射自显影和化学摄影。该设施的服务被广泛用于评估蛋白质纯化、细胞免疫沉淀、重组蛋白的表达以及来自培养细胞、组织和重组宿主的免疫印迹。这些技术和方法对于我们研究我们所使用的基本上所有基因和蛋白质的表达和纯化都是至关重要的。此外，还可以确定 在转基因和基因敲除小鼠中，通过免疫印迹研究的基因的组织特异性表达模式严重依赖于Core的这一方面。

The Molecular Biology Core Laboratory is directed by Dr. Russell. He will be assisted in day-to-day operations by Dr. Jonathan Cohen, who is an expert in high-throughput DMA sequencing. This Core laboratory provides support for acrylamide gel electrophoresis of proteins, DMA sequencing, analysis of gene expression by oligonucleotide microarray hybridization and real-time polymerase chain reactions (PCRs), oligonucleotide procurement, genomic DMA and RNA isolation, and the maintenance and storage of bacterial strains, plasmids, and purified proteins used within this Program Project. Four experienced technicians, Kevin Anderson (100% time), Emily Brown (100%), Jeffrey Cormier (100%), and Scott Clark (25%) will perform the duties associated with this Core. The laboratory facility is located within the Department of Molecular Genetics. For the analysis of proteins, 30 vertical electrophoresis units and 12 multi-outlet power supplies are available. SDS polyacrylamide gels are either prepared (25% of gels) or purchased pre-poured (75% of gels) and run by a single technician (Emily Brown). Following electrophoresis, individual Investigators working on the different Research Projects process the gels for autoradiography, immunoblotting, or protein sequencing. A darkroom that contains a Konica automatic X-ray developer is used to process all autoradiograms and chemilumigrams. The services of the facility are used extensively to assess protein purification, immunoprecipitation from cells, expression of recombinant proteins, and immunoblotting from cultured cells, tissues, and recombinant hosts. These techniques and methods are crucial to our studies on the expression and purification of essentially all genes and proteins with which we work. In addition, the determination of tissue-specific expression patterns of genes under study via immunoblotting in transgenic and knockout mice is heavily dependent on this aspect of the Core.